| Background:At present,cardiovascular and cerebrovascular diseases are still the first cause of human death,which is a serious threat to human health.Arterosclerosis(AS)is the early pathological basis of most cardiovascular and cerebrovascular diseases,and it is a complex chronic progressive inflammation.Disease[1]can affect systemic symptoms,in which the liver acts as an important metabolic organ and systemic systemic inflammatory response center,and various inflammatory factors and body fluid production organs and target organs.These inflammatory mediators play a synergistic and antagonistic role in the initiation and progression of AS-related diseases[2]so the liver is one of the most vulnerable organs in the formation of AS.Combined with this experiment,it was found in the nature of the previous period that the liver of the AS model group showed a significant decrease in the volume of the liver,which was pale yellow,which was adherent to the surrounding tissues,and the pathological morphology and abnormal liver function of the cut surface were greasy and dull..We conducted a related literature review and found that liver damage is closely related to AS.Hepatocyte adipose disease and its accompanying hepatic necroinflammation have a promoting effect on AS[3].Therefore,studying the pathological state of the liver during the pathogenesis of AS has guiding significance for the prevention and treatment of AS.This study used the right carotid collar placement(PCCP)combined with high-fat diet formula to construct the AS model,taking the liver injury as the entry point during the formation of AS,and observing the Simiao Yongan Decoction and the liver.The injury is closely related to the intervention of PPAR y and its related inflammatory signaling pathway,and the protective mechanism of Simiao Yongan Decoction on AS liver injury is discussed.Methods:Experiment 1:Compare the effects of different modeling methods on the construction of AS model in ApoE-/-mice.The AS model was constructed using a high-fat diet combined with a PCCP method.There were 4 groups in this experiment:C57 group,high fat group,sham operation group and model group.The mice in each group were fed for 3 days.Except the C57 group,the other groups were fed with high fat.Two weeks after high-fat feeding,the model group underwent PCCP surgery.The model of AS model was observed by small animal ultrasound at 3d,4w and 8w after operation.The other groups except the C57 group continued high-fat feeding for 8 weeks.After taking the materials,the blood lipids were selected(total cholesterol(TC),triacylglyceride(TG),high density lipoprotein cholesterol(HDL-C)and low density lipoprotein cholesterol,(low density lipoprotein cholesterol,LDL-C))and liver function alanine aminotransferase(ALT)and aspartate aminotransferase(AST)determination;HE staining and Masson staining to observe the pathological morphology of the right common carotid artery.The above indicators were used to evaluate the effects of different modeling methods on the construction of AS models in ApoE-/-mice.Experiment 2:Pharmacodynamics and molecular mechanism of Simiao Yongan Decoction on inflammatory response of liver injury in ApoE-/-mice AS model.The AS model was constructed by high-fat diet combined with PCCP method and divided into 5 groups:sham operation group,model group,atorvastatin group,pioglitazone group and Simiao Yongan Decoction group.Three treatment groups were intragastrically administered,and the sham operation group and the model group were given an equal volume of deionized water.All groups of mice were fed with high fat throughout the course and were taken 8 weeks after surgery.The biochemical analyzer was used to detect the content of four lipids and AST and ALT.HE staining and oil red O staining were used to observe the pathological morphology of liver tissue.Multi-factor detection technique was used to detect interleukin-6(IL-6)in serum.Interleukin-17A(IL-17A),interferon-?(interferon-?,IFN ?)inflammatory cytokine expression.Immunohistochemistry was used to detect peroxisome proliferator-activated receptor gamma(PPAR ?)and cytokine signaling inhibitor 1(SOCS1).Protein level expression of programmed cell death 4(PDCD4)and nuclear transcription factor-?B(NF-?B);Western blot analysis of liver PPAR ?,SOCS1,PDCD4 in each group of mice,NF-? Bp65,NLRP3,phosphorylated signal of phosphatylated signal transducer and activator of transcription 3(p-STAT3)Horizontal expression.Results:Experiment one(1)Ultrasound results showed that the blood flow of the common carotid artery and internal carotid artery was unobstructed three days after surgery,suggesting that the operation was successful.The intimal-medial thickness(IMT)of the common carotid artery in the model group was gradually thickened,the vascular lumen was further narrowed,and the peak systolic velocity(PSV)and mean of the internal carotid artery were averaged.The mean velocity(MV)is gradually increased.(2)Four items of blood lipids showed that compared with C57 group,the levels of TC,TG and LDL-C in high-fat group were significantly increased(P<0.01,P<0.05,P<0.01),and HDL-C was significantly decreased(P<0.01).Compared with the sham operation group,the HDL-C of the model group was significantly increased(P<0.01),and the TC,TG,and LDL-C were not statistically different(P>0.05).Compared with the high fat group,the TG and LDL of the model group were compared.-C decreased,HDL-C increased significantly(P<0.05,P<0.01,P<0.05),and there was no statistical difference in TC(P>0.05).(3)Liver function indicators showed that there was no significant difference in AST and ALT levels between the high-fat group and the C57 group(P>0.05).Compared with the sham-operated group,the model group had significantly increased AST and ALT(P<0.01,P<0.01);compared with the high-fat group,the AST and ALT of the model group were significantly increased,the difference was statistically significant(P<0.01,P<0.01).(4)Carotid pathology showed that the common carotid artery of C57 group was intact,the thickness was uniform,and the VSMC of the middle membrane was neatly arranged.No lesions of AS lesions were observed.Compared with the C57 group,the wall thickness of the carotid wall was uneven.Membrane thickening is not obvious,a small amount of foam cells are aggregated,which is typical AS early lesion-lipid streak stage;sham operation group has thin inner thickness and uneven inner membrane,compared with sham operation,model group mice The membrane was thickened obviously,a large number of plaques formed,and a large amount of collagen fibers and inflammatory cells infiltrated in the plaque,which belonged to the late stage of AS-fiber atherosclerotic plaque.Compared with the high-fat group,the AS lesions in the model group were significantly aggravated.The blood vessel wall protrudes into the lumen and is filled with a large number of lipid foam cells.Masson staining showed that compared with the model group,only a small amount of collagen fibers were seen in the other three groups of carotid lumens,and AS lesions were significantly alleviated.Experiment 2(1)Four blood lipids and liver function indicators showed that serum HDL-C was significantly increased in the model group compared with the sham operation group.Compared with the model group,HDL-C was significantly decreased and LDL-C was significantly increased in each drug-administered group.There was no significant change in TC and TG between the groups,and the difference was not statistically significant.Compared with the sham operation group,serum ALT and AST in the model group were significantly increased.Compared with the model group,AST and ALT were significantly decreased in each group,and the difference was statistically significant.(2)Histopathological observation of the liver showed that the lobular structure of the sham operation group was basically normal;compared with the sham operation group,the hepatic sinus was narrowed in the model group,and spotted or focal hepatocyte necrosis was observed in the lobule with a large amount of inflammatory cell infiltration.After the intervention,the liver injury was significantly reduced compared with the model group,and no obvious hepatocyte necrosis and inflammatory cell infiltration were observed.Oil red O staining showed that the liver of each group showed lipid deposition,but there was no significant difference between the groups.(3)The results of inflammatory factors showed that compared with the sham operation group,the expression of IL-6,IL-17A and IFN ? in the model group was significantly increased(P<0.01,P<0.05,P<0.05);compared with the model group,Simiao Yongan Decoction group can significantly reduce the expression of IL-6,IL-17A and IFN ? in serum(P<0.05),and its effect is consistent with atorvastatin and pioglitazone.(4)The results of immunohistochemistry showed that compared with the sham operation group,the positive expression of PPAR y and SOCS1 in the model group decreased,and the positive expression of PDCD4 and NF-?B increased.Compared with the model group,the atorvastatin group,the pioglitazone group and the Simiao Yongan Decoction group PPAR y and SOCS1 positive expression increased,and PDCD4 and NF-?B positive expression decreased.(5)Western Blot results showed that compared with the sham operation group,the expression of PPAR y and SOCS1 protein in the model group was significantly decreased(P<0.01),and the expression levels of p-JAK2,p-STAT3,PDCD4,NF-?B p65 and NLRP3 were significantly increased(P<0.01);compared with the model group,PPAR y and SOCS1 protein expression levels were significantly increased in the atorvastatin group,pioglitazone group and Simiao Yongan Decoction group(P<0.01),while p-JAK2,p-STAT3 The expression levels of PDCD4,NF-?B p65 and NLRP3 protein were significantly lower,which was statistically significant(P<0.01).Conclusions:1 Compared with the traditional high-fat feeding AS model,the high-fat diet combined with the PCCP method can shorten the experimental period and successfully construct the AS model quickly.2 Simiao Yongan Decoction can improve liver tissue damage by inhibiting the expression of NLRP-3,IL-6,IL-17A and IFN ?3 Simiao Yongan Decoction may improve inflammatory response of ApoE-/-mice with liver injury by regulating NF-?B and JAK2/STAT3 signaling pathway through PPAR?... |
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